Deflamin Attenuated Lung Tissue Damage in an Ozone-Induced COPD Murine Model by Regulating MMP-9 Catalytic Activity.

Chronic obstructive pulmonary disease (COPD) is comprised of histopathological alterations such as pulmonary emphysema and peribronchial fibrosis. Matrix metalloproteinase 9 (MMP-9) is one of the key enzymes involved in both types of tissue remodeling during the development of lung damage. In recent studies, it was demonstrated that deflamin, a protein component extracted from Lupinus albus, markedly inhibits the catalytic activity of MMP-9 in experimental models of colon adenocarcinoma and ulcerative colitis. Therefore, in the present study, we investigated for the first time the biological effect of deflamin in a murine COPD model induced by chronic exposure to ozone. Ozone exposure was carried out in C57BL/6 mice twice a week for six weeks for 3 h each time, and the treated group was orally administered deflamin (20 mg/kg body weight) after each ozone exposure. The histological results showed that deflamin attenuated pulmonary emphysema and peribronchial fibrosis, as evidenced by H&E and Masson's trichrome staining. Furthermore, deflamin administration significantly decreased MMP-9 activity, as assessed by fluorogenic substrate assay and gelatin zymography. Interestingly, bioinformatic analysis reveals a plausible interaction between deflamin and MMP-9. Collectively, our findings demonstrate the therapeutic potential of deflamin in a COPD murine model, and suggest that the attenuation of the development of lung tissue damage occurs by deflamin-regulated MMP-9 catalytic activity.

Comparative Screening of the Liver Gene Expression Profiles from Type 1 and Type 2 Diabetes Rat Models.

Experimental animal models of diabetes can be useful for identifying novel targets related to disease, for understanding its physiopathology, and for evaluating emerging antidiabetic treatments. This study aimed to characterize two rat diabetes models: HFD + STZ, a high-fat diet (60% fat) combined with streptozotocin administration (STZ, 35 mg/kg BW), and a model with a single STZ dose (65 mg/kg BW) in comparison with healthy rats. HFD + STZ- induced animals demonstrated a stable hyperglycemia range (350-450 mg/dL), whereas in the STZ-induced rats, we found glucose concentration values with a greater dispersion, ranging from 270 to 510 mg/dL. Moreover, in the HFD + STZ group, the AUC value of the insulin tolerance test (ITT) was found to be remarkably augmented by 6.2-fold higher than in healthy animals (33,687.0 ± 1705.7 mg/dL/min vs. 5469.0 ± 267.6, respectively), indicating insulin resistance (IR). In contrast, a more moderate AUC value was observed in the STZ group (19,059.0 ± 3037.4 mg/dL/min) resulting in a value 2.5-fold higher than the average exhibited by the control group. After microarray experiments on liver tissue from all animals, we analyzed genes exhibiting a fold change value in gene expression <-2 or >2 (p-value <0.05). We found 27,686 differentially expressed genes (DEG), identified the top 10 DEGs and detected 849 coding genes that exhibited opposite expression patterns between both diabetes models (491 upregulated genes in the STZ model and 358 upregulated genes in HFD + STZ animals). Finally, we performed an enrichment analysis of the 849 selected genes. Whereas in the STZ model we found cellular pathways related to lipid biosynthesis and metabolism, in the HFD + STZ model we identified pathways related to immunometabolism. Some phenotypic differences observed in the models could be explained by transcriptomic results; however, further studies are needed to corroborate these findings. Our data confirm that the STZ and the HFD + STZ models are reliable experimental models for human T1D and T2D, respectively. These results also provide insight into alterations in the expression of specific liver genes and could be utilized in future studies focusing on diabetes complications associated with impaired liver function.

Combined Gamma Conglutin and Lupanine Treatment Exhibits In Vivo an Enhanced Antidiabetic Effect by Modulating the Liver Gene Expression Profile.

Previous studies have individually shown the antidiabetic potential of gamma conglutin (Cγ) and lupanine from lupins. Until now, the influence of combining both compounds and the effective dose of the combination have not been assessed. Moreover, the resulting gene expression profile from this novel combination remains to be explored. Therefore, we aimed to evaluate different dose combinations of Cγ and lupanine by the oral glucose tolerance test (OGTT) to identify the higher antidiabetic effect on a T2D rat model. Later, we administered the selected dose combination during a week. Lastly, we evaluated biochemical parameters and liver gene expression profile using DNA microarrays and bioinformatic analysis. We found that the combination of 28 mg/kg BW Cγ + 20 mg/kg BW lupanine significantly reduced glycemia and lipid levels. Moreover, this treatment positively influenced the expression of Pdk4, G6pc, Foxo1, Foxo3, Ppargc1a, Serpine1, Myc, Slc37a4, Irs2, and Igfbp1 genes. The biological processes associated with these genes are oxidative stress, apoptosis regulation, and glucose and fatty-acid homeostasis. For the first time, we report the beneficial in vivo effect of the combination of two functional lupin compounds. Nevertheless, further studies are needed to investigate the pharmacokinetics and pharmacodynamics of the Cγ + lupanine combined treatment.

[Metabolic profile and concentration of ghrelin and obestatin in children and adolescents with obesity]. / Perfil metabólico, concentración de grelina y obestatina en niños y adolescentes con obesidad.

Background: It has been pointed out that ghrelin and obestatin could have an impact on the genesis of obesity, since they estimulate and inhibit apetite and, therefore, food consumption. Objective: To compare the metabolic profile, lipid profile and the concentrations of ghrelin and obestatin in children with normal weight or obesity. Material and methods: Cross-sectional design with 97 normal weight or obese children, 6 to 18 years of age, who did not present systemic diseases. The serum concentrations of glucose, insulin, total cholesterol, triglycerides, high (HDL), low (LDL) and very low density (VLDL) lipoproteins, aspartate aminotransferase (AST), alanine aminotransferase (ALT), ghrelin and obestatin were determined. Descriptive statistics were performed. Student's t test was used to compare groups, and correlation coefficients of ghrelin and obestatin values with biochemical and anthropometric variables. A p value of ≤ 0.05 was significant. Results: 55 children with normal weight and 42 with obesity were included; mean age was 10.7 years. Triglycerides, LDL, VLDL, ALT and insulin were higher, and HDL lower in obese children (p < 0.05). Ghrelin values were higher in normal weight children (p < 0.05), and there was no difference in obestatin values. Conclusions: The lower concentration of ghrelin in obese children may indicate a negative feedback to regulate energy consumption. Children and adolescents with obesity show metabolic and lipid profile alterations that place them at risk of early development of cardiovascular risk factors.

Introducción: se ha señalado que la grelina y la obestatina podrían incidir en la génesis de la obesidad al estimular o inhibir el apetito y, por ende, el consumo de alimentos. Objetivo: comparar el perfil metabólico, el perfil de lípidos y las concentraciones de grelina y obestatina en niños con normopeso u obesidad. Material y métodos: diseño transversal con 97 niños de 6 a 18 años con normopeso u obesidad que no presentaran enfermedades sistémicas. Se determinaron las concentraciones séricas de glucosa, insulina, colesterol total, triglicéridos, lipoproteínas de colesterol de alta (HDL), baja (LDL) y muy baja densidad (VLDL), aspartato aminotransferasa (AST), alanina aminotransferasa (ALT), grelina y obestatina. Se usó estadística descriptiva. Se utilizó la prueba t de Student para comparar grupos, y coeficientes de correlación de los valores de grelina y obestatina con las variables bioquímicas y antropométricas. Un valor de p ≤ 0.05 fue significativo. Resultados: se incluyeron 55 niños con normopeso y 42 con obesidad; la edad promedio fue de 10.7 años. Los triglicéridos, LDL, VLDL, ALT y la insulina fueron superiores, y el HDL inferior en niños con obesidad (p < 0.05). Los valores de la grelina fueron superiores en niños con normopeso (p < 0.05) y no hubo diferencia en los de la obestatina. Conclusiones: la menor concentración de grelina en niños con obesidad puede indicar una retroalimentación negativa para regular el consumo de energía. Los niños y adolescentes con obesidad muestran alteraciones metabólicas y del perfil de lípidos que los ponen en riesgo de desarrollar tempranamente factores de riesgo cardiovascular.

Nanoparticles Formulation Improves the Antifibrogenic Effect of Quercetin on an Adenine-Induced Model of Chronic Kidney Disease.

Renal fibrosis is the final stage of chronic kidney injury characterized by glomerulosclerosis and tubulointerstitial fibrosis with parenchymal destruction. Quercetin belongs to the most studied flavonoids with antioxidant, anti-inflammatory, antifibrogenic, and antitumor activity. It modifies the TGF-ß/Smad signaling pathway, decreasing profibrogenic expression molecules and inducing the expression of antioxidant, anti-inflammatory, and antifibrogenic molecules. However, quercetin exhibits poor water solubility and low absorption and bioavailability. This limitation was solved by developing a nanoparticles formulation that improves the solubility and bioavailability of several bioactive compounds. Therefore, we aimed to investigate the in vivo antifibrogenic effect of a quercetin nanoparticles formulation. Male C57BL/6 mice were induced into chronic renal failure with 50 mg/kg of adenine for four weeks. The animals were randomly grouped and treated with 25, 50, or 100 mg/kg of quercetin, either macroparticles or nanoparticles formulation. We performed biochemical, histological, and molecular analyses to evaluate and compare the effect of macroparticles versus nanoparticles formulation on kidney damage. Here, we demonstrated that smaller doses of nanoparticles exhibited the same beneficial effect as larger doses of macroparticles on preventing kidney damage. This finding translates into less quercetin consumption reaching the desired therapeutic effect.

Cladodes from Nopalea cochenillifera (L.) Salm-Dyck (Cactaceae) attenuate postprandial glycaemia without markedly influencing α-glucosidase activity.

Although the cladodes of Nopalea cochenillifera are used in Mexican traditional medicine to treat diabetes and various other diseases, its antihyperglycaemic properties, phenolic content, and antioxidant activity are not well documented. Thus, we determined the activity of a single dose of fresh, blended cladodes on postprandial glycaemia in rats after a starch load. We prepared a methanolic extract of N. cochenillifera cladodes and measured its phenolic content, antioxidant capacity, and α-glucosidase inhibitory activity. The antihyperglycaemic effect of blended cladodes was similar to that of acarbose when considering the changes in glucose levels from baseline. Furthermore, the methanolic extract contained a considerable amount of phenolic compounds and exhibited antioxidant activity in the DPPH assay, but did not markedly inhibit α-glucosidase and had a low antioxidant effect in the ABTS test.

Lupin protein isolate improves insulin sensitivity and steatohepatitis in vivo and modulates the expression of the Fasn, Gys2, and Gsk3b genes.

Although studies on lupin protein isolate (LPI) have indicated the presence of a preventive effect on insulin resistance (IR) and lipid disturbances, their influence on established pathological traits has received little attention. Here, we evaluated the in vivo effects of LPI on IR and steatohepatitis as well as its influence on genes involved in lipid and carbohydrate metabolism. We first induced IR and steatohepatitis in rats by maintaining them on a high-fat diet for 5 weeks. Thereafter, we administered LPI to the rats daily for 3 weeks. LPI improved insulin sensitivity (AUC: 6,777 ± 232 vs. 4,971 ± 379, p < .05, pre- vs. post-treatment values) and reduced glucose and triglyceride levels by one-third. In addition, LPI-treated rats exhibited attenuated steatohepatitis. At the molecular level, LPI treatment reduced liver Fasn gene expression substantially but increased Gys2 and Gsk3b levels. We concluded that the hypolipidemic and hypoglycemic activities of LPI may be caused by reduced liver lipogenesis and modulation of insulin sensitization mechanisms.

Effect of Lupinus rotundiflorus gamma conglutin treatment on JNK1 gene expression and protein activation in a rat model of type 2 diabetes.

CONTEXT: Gamma conglutin (Cγ) from lupine species represents a potential complementary treatment for type 2 diabetes mellitus (T2DM) because of its hypoglycaemic effect. However, its underlying mechanism of action is not fully known. OBJECTIVE: To evaluate whether Cγ from Lupinus rotundiflorus M. E. Jones (Fabaceae) modulates c-Jun N-terminal kinase 1 (JNK1) expression and activation in a T2DM rat model. MATERIALS AND METHODS: Gamma conglutin isolated from L. rotundiflorus seeds was characterized by SDS-PAGE. Fifteen Wistar rats with streptozotocin-induced T2DM (HG) were randomized into three groups (n = 5): vehicle administration (HG-Ctrl), oral treatment with Cγ (120 mg/kg/day) (HG-Lr) for one week, and treatment with metformin (300 mg/kg/day) (HG-Met); a healthy group (Ctrl, n = 5) was included as control. The levels of glucose and biomarkers of renal and hepatic function were measured pre- and post-treatment. Hepatic Jnk1 expression and phosphorylation of JNK1 were evaluated by qRT-PCR and western blot, respectively. RESULTS: Oral treatment with either Cγ or metformin reduced serum glucose level to 86.30 and 74.80 mg/dL, respectively (p ˂ 0.05), from the basal levels. Jnk1 expression was 0.65- and 0.54-fold lower (p ˂ 0.05) in the HG-Lr and HG-Met groups, respectively, than in HG-Ctrl. Treatment with Cγ decreased JNK1 phosphorylation. However, Cγ did not change the levels of kidney and liver biomarkers. DISCUSSION AND CONCLUSIONS: Treatment with Cγ from L. rotundiflorus inhibited Jnk1 expression, in vivo, suggesting JNK1 as a potential therapeutic target in diabetes and revealing one mechanism underlying the hypoglycaemic effect of lupine Cγ. Nevertheless, further studies are required.

Analysis of hepatic transcriptome modulation exerted by γ-conglutin from lupins in a streptozotocin-induced diabetes model.

Lupinus albus γ-conglutin is proposed to positively affect glucose metabolism through inhibition of hepatic glucose production and insulin-mimetic activity; however, the action mechanism is not entirely known. Besides, most studies had focused on its effect on molecular targets directly related to glucose metabolism, and few studies have investigated how γ-conglutin may affect the liver gene expression or if it plays a role in other metabolic processes. Therefore, we investigated the influence of γ-conglutin on the liver transcriptome of streptozotocin-induced diabetic rats using DNA microarrays, ontological analyses, and quantitative PCR. Of the 22,000 genes evaluated, 803 and 173 were downregulated and upregulated, respectively. The ontological analyses of the differentially expressed genes revealed that among others, the mitochondria, microtubules, cytoskeleton, and oxidoreductase activity terms were enriched, implying a possible role of γ-conglutin on autophagy. To corroborate the microarray results, we selected and quantified, by PCR, the expression of two genes associated with autophagy (Atg7 and Snx18) and found their expression augmented two and threefold, respectively; indicating a higher autophagy activity in animals treated with γ-conglutin. Although complementary studies are required, our findings indicate for the first time that the hypoglycaemic effects of γ-conglutin may involve an autophagy induction mechanism, a pivotal process for the preservation of cell physiology and glucose homeostasis.

[Glucotransporters: clinical, molecular and genetic aspects]. / Glucotransportadores (GLUT): Aspectos clínicos, moleculares y genéticos.

Oxidation of glucose is the major source of obtaining cell energy, this process requires glucose transport into the cell. However, cell membranes are not permeable to polar molecules such as glucose; therefore its internalization is accomplished by transporter proteins coupled to the cell membrane. In eukaryotic cells, there are two types of carriers coupled to the membrane: 1) cotransporter Na+-glucose (SGLT) where Na+ ion provides motive power for the glucose´s internalization, and 2) the glucotransporters (GLUT) act by facilitated diffusion. This review will focus on the 14 GLUT so far described. Despite the structural homology of GLUT, different genetic alterations of each GLUT cause specific clinical entities. Therefore, the aim of this review is to gather the molecular and biochemical available information of each GLUT as well as the particular syndromes and pathologies related with GLUT´s alterations and their clinical approaches.

A cut-point value of uncarboxylated to carboxylated index is associated with glycemic status markers in type 2 diabetes.

BACKGROUND: The uncarboxylated osteocalcin (ucOC) has been described as a regulator of glucose metabolism in mice, and it is decreased in human type 2 diabetes mellitus (T2D). Although inversely correlated with serum glucose, insulin, and glycated hemoglobin, it is unclear if ucOC decrement is caused by diabetes or plays a role in the pathogenesis and/or progression of the disease. Whatever the case may be, diabetes affects osteoblast gene expression, and possibly the proportion of ucOC over carboxylated OC (cOC). The association of ucOC/cOC index with glycemic status markers in patients with T2D has not been described before. OBJECTIVE: The objective of this study was to assess the ucOC/cOC index and its relationship with glycemic status markers in patients with T2D. METHODS: The ucOC/cOC index was determined by the quotient of ucOC and cOC serum levels in 80 T2D patients and 160 healthy subjects. The relationship between the ucOC/cOC index and glycemic status markers was evaluated. RESULTS: The ucOC/cOC index was low and negatively correlated to fasting plasma glucose and homeostasis assessment-insulin resistance model in T2D patients. The odds ratio for T2D patients with an ucOC/cOC index below the cut-point obtained by receiver operating characteristic analysis was 12.64 (confidence interval, 5.75-27.77; P < 0.001). CONCLUSIONS: A value of ucOC/cOC index less than 0.3 is associated with markers of poor metabolic control in patients with T2D.

Inhibitory mechanisms of two Uncaria tomentosa extracts affecting the Wnt-signaling pathway.

Uncaria tomentosa ("uña de gato"; "cat's claw"), a woody vine native to the Amazon rainforest, is commonly used in South American traditional medicine to treat a broad spectrum of diseases. Although recent studies have reported anti-inflammatory and anti-proliferative properties of different alkaloids extracted from this plant, the underlying molecular mechanisms of these effects have not been elucidated yet. Our study investigates the inhibitory mechanisms of Uncaria tomentosa extracts on the Wnt-signaling pathway, a central regulator of development and tissue homoeostasis. A modified cell-based luciferase assay for screening inhibitors of the Wnt-pathway was used for analysis. Three cancer cell lines displaying different levels of aberrant Wnt-signaling activity were transfected with Wnt-signaling responsive Tcf-reporter plasmids and treated with increasing concentrations of two Uncaria tomentosa bark extracts. Wnt-signaling activity was assessed by luciferase activity and by expression of Wnt-responsive target genes. We show that both, an aqueous and an alkaloid-enriched extract specifically inhibit Wnt-signaling activity in HeLa, HCT116 and SW480 cancer cells resulting in reduced expression of the Wnt-target gene: c-Myc. The alkaloid-enriched extract (B/S(rt)) was found to be more effective than the aqueous extract (B/W(37)). The strongest effect was observed in SW480 cells, displaying the highest endogenous Wnt-signaling activity. Downregulation of Wnt-signaling by a dominant negative-TCF-4 variant in non-cancer cells rendered the cells insensitive towards treatment with B/S(rt). B/Srt was less toxic in non-cancer cells than in cancer cells. Our data suggest that the broad spectrum of pharmacological action of Uncaria tomentosa involves inhibition of the Wnt-signaling pathway, downstream of beta-Catenin activity.

[Lung cancer histological types and diagnostic methods in a tertiary care facility]. / Tipos histológicos y métodos diagnósticos en cáncer pulmonar en un centro hospitalario de tercer nivel.

OBJECTIVE: To describe the histological frequencies, diagnostic methods, gender and age of lung cancer patients in a third level care hospital in western Mexico. METHODS: A retrospective and descriptive study (2002 to 2005) was undertaken. From all reviewed cases, 242 patients had a diagnosis compatible with lung cancer. We analyzed the histological type, gender, age group, and diagnostic method. RESULTS: The histological type frequencies of lung cancer were as follows: 63.6% adenocarcinoma, 19.0% squamous cell carcinoma, 15.2% small cell carcinoma, and 2.2% large-cell carcinoma. Among these patients, a male-female ratio of 2:1 with a mean age of 63 years was found. Regarding the diagnostic methods followed, the most common included: transthoracic needle aspiration, bronchial biopsy, and bronchial washing. CONCLUSIONS: Adenocarcinoma was the most frequent histological type in the sample studied, an association between female gender and risk of developing adenocarcinoma was detected.

Tipos histológicos y métodos diagnósticos en cáncer pulmonar en un centro hospitalario de tercer nivel / Lung cancer histological types and diagnostic methods in a tertiary care facility

Objetivo: Describir las frecuencias de tipos histológicos, métodos diagnósticos, sexo y edad de pacientes con cáncer pulmonar, en el Centro Médico Nacional de Occidente del Instituto Mexicano del Seguro Social, Guadalajara, Jalisco. Métodos: Estudio descriptivo y retrospectivo de los casos revisados entre 2002 y 2005; 242 pacientes presentaron diagnóstico de cáncer pulmonar. Se analizó el tipo histológico, sexo, edad y métodos diagnósticos. Resultados: Los tipos histológicos de cáncer pulmonar presentaron las siguientes frecuencias: adenocarcinoma 63.6%, carcinoma de células escamosas 19.0%, carcinoma de células pequeñas 15.2% y carcinoma de células grandes 2.2%. Se encontró una relación varón:mujer de 2:1, con una edad promedio de 63 años. Los métodos diagnósticos más utilizados fueron biopsia por aspiración con aguja fina, biopsia bronquial y lavado bronquial. Conclusiones: En la población analizada, el adenocarcinoma fue el tipo histológico más frecuente. Además, se encontró asociación entre el sexo femenino y el riesgo de presentar adenocarcinoma.

OBJECTIVE: To describe the histological frequencies, diagnostic methods, gender and age of lung cancer patients in a third level care hospital in western Mexico. METHODS: A retrospective and descriptive study (2002 to 2005) was undertaken. From all reviewed cases, 242 patients had a diagnosis compatible with lung cancer. We analyzed the histological type, gender, age group, and diagnostic method. RESULTS: The histological type frequencies of lung cancer were as follows: 63.6% adenocarcinoma, 19.0% squamous cell carcinoma, 15.2% small cell carcinoma, and 2.2% large-cell carcinoma. Among these patients, a male-female ratio of 2:1 with a mean age of 63 years was found. Regarding the diagnostic methods followed, the most common included: transthoracic needle aspiration, bronchial biopsy, and bronchial washing. CONCLUSIONS: Adenocarcinoma was the most frequent histological type in the sample studied, an association between female gender and risk of developing adenocarcinoma was detected.